Knock-down of apoptosis inducing factor gene protects endoplasmic reticulum stress-mediated goat granulosa cell apoptosis

Abstract

The apoptosis of granulosa cells is the main cause of follicular atresia, and endoplasmic reticulum (ER) stress is involved in the apoptosis of granulosa cells. Apoptosis inducing factor (AIF) mediates caspase-independent apoptosis and causes chromatin condensation and DNA fragmentation, but its role in ER stress-mediated granulosa cell apoptosis during goat follicular atresia remains largely unknown. The aim of this study was to investigate the function of AIF in the apoptosis of goat granulosa cells mediated by ER stress. The results of immunohistochemical and Western blot analyses demonstrated that AIF was mainly located in granulosa cells, and the expression of AIF significantly increased during follicular atresia. Then, AIF-short hairpin RNA recombinant lentiviral vectors were constructed successfully and transfected into human telomerase reverse transcriptase-goat granulosa cells (hTERT-GGCs). Real-time quantitative polymerase chain reaction and Western blot analysis confirmed that AIF was effectively knocked down in hTERT-GGCs. Flow cytometry results showed that the knockdown of AIF in hTERT-GGCs reduced apoptosis due to serum starvation or thapsigargin (Tg) treatment. In addition, AIF depletion changed the expression of related molecular marker molecules of ER stress under Tg treatment. In conclusion, AIF may serve as a key factor during follicular atresia, and AIF depletion protects ER stress-mediated goat granulosa cell apoptosis.

Keywords: Caspase-independent death effector; Follicular atresia; Short hairpin interfering RNA; Unfolded protein response.